急性髓系白血病新型预后分子标志物NKX2-3的发现

Discovery of A New Prognostic Molecular Marker NKX2-3 for Acute Myeloid Leukemia

【目的】通过分析生物信息数据库中影响急性髓系白血病(AML)患者的预后的分子标志物的表达,为进一步探索AML预后的新型分子标志物提供实验基础。【方法】从癌症基因组图谱(TCGA)生物信息数据库中的179例AML患者的预后数据进行差异性基因分析及生存分析;利用基因表达集锦(GEO)数据库中的GSE13159数据集的74例健康人(HI)骨髓标本与542例AML初发患者骨髓标本进行分析,检测差异性目的基因表达水平在AML初发患者与健康人中的差异性;收集初发18例AML患者的外周血及骨髓样本,同时收集年龄和性别匹配的20例健康志愿者的样本作为对照,采用实时荧光定量PCR验证差异基因在AML患者体内的表达水平。【结果】生物信息数据分析显示,根据R语言计算出的NK2转录因子相关基因位点3(NKX2-3)的最佳截断值0.051进行生存分析,发现与低表达NKX2-3的AML初发患者相比,高表达NKX2-3的AML初发患者总体生存率较差(P=0.0036);与HI相比,NKX2-3在AML初发组患者中显著高表达(P<0.001);实时荧光定量PCR的验证结果也证实NKX2-3-1和NKX2-3-2在AML初发组患者中高表达,且与健康人组相比有显著相关性(P=0.000,P=0.000)。【结论】NKX2-3在AML初发组患者中高表达,且高表达NKX2-3的AML患者总体生存较差;NKX2-3可能与AML临床转归与预后密切相关。

【Objective】To analyze the expression of molecular marker affecting the prognosis of acute myeloid leukemia(AML)patients from bioinformatics database,thus providing an experimental basis for further exploration of a novel molecular marker for the prognosis of AML.【Methods】The prognostic data of 179 AML patients from The Cancer Genome Atlas(TCGA)database were examined for differential gene analysis and survival analysis.The bone marrow samples of 74 healthy individuals(HI)and 542 de novo AML patients in the dataset GSE13159 downloaded from the Gene Expression Omnibus(GEO)database were analyzed to detect the difference in the expression levels of differential target genes.Peripheral blood and bone marrow samples were collected from 18 de novo AML patients and 20 age-and gender-matched healthy controls,and real-time fluorescent quantitative PCR was used to validate the expression levels of the differential genes in the AML patients.【Results】Bioinformatics data analysis showed that the optimal cut-off value of Homo sapiens NK2 homeobox 3(NKX2-3)calculated by R language was 0.051.Survival analysis revealed a statistically poorer overall survival in de novo AML patients with high NKX2-3 expression than in those with low NKX2-3 expression(P=0.0036).NKX2-3 was highly expressed in patients with de novo AML than in HI and the difference was statistically significant(P<0.001).Real-time fluorescence quantitative PCR verified the expression levels of the NKX2-3 gene in AML patients and confirmed that compared with those in HI,in the de novo AML patients,NKX2-3-1 and NKX2-3-2 were highly expressed and were significantly correlated(P=0.000,P=0.000).【Conclusion】NKX2-3 is highly expressed in de novo AML patients,and the AML patients with high NKX2-3 expression have poor overal survival.NKX2-3 may be closely related to the clinical outcome and prognosis of AML.